Characterisation of protein kinase C like activities in Trypanosoma brucei.
Protein kinase activities in bloodstream and procyclic forms of Trypanosoma brucei have been partially purified and characterised. Cytosolic extracts were separated on DEAE-cellulose and assayed for the ability to phosphorylate histone in the presence of Ca2+ and diacylglycerol. Five peaks of activity were identified in bloodstream T. brucei and only three in procyclic lysates. One of the kinases present in bloodstream T. brucei shares may characteristics with mammalian protein kinase C. Further characterisation of the kinases using an in situ assay after separating proteins by isoelectric focussing confirmed that the kinases present in bloodstream and procyclic stages differed in properties and either bloodstream kinases are more stable or greater in number than procyclic kinases. A protein present in bloodstream T. brucei was shown by Western blot analysis to contain an epitope recognized by a monoclonal antibody raised against mammalian protein kinase C. We thus conclude that the protein kinases are differentially regulated between the two stages of the parasite and that the bloodstream stage has a protein kinase C homologue. The implications of these findings in relation to a cellular signalling pathway in trypanosomes is discussed.
Keith
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Wellcome Unit of Molecular Parasitology, University of Glasgow, Scotland, U.K.
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Tait
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D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002118:Calcium / Q000494:pharmacology; D005591:Chemical Fractionation; D002848:Chromatography, DEAE-Cellulose; D004075:Diglycerides / Q000494:pharmacology; D006657:Histones / Q000378:metabolism; D007526:Isoelectric Point; D010766:Phosphorylation; D011493:Protein Kinase C / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D004075:Diglycerides; D006657:Histones; D011493:Protein Kinase C; D002118:Calcium